Sulphonamide compound in oil



April 22, 1941. n. R. cLlMl-:NKO 2,238,973

SULPHONAMIDE COMPOUND IN OIL Filed July 26, 1940 5 Sheets-Sheet 2 P4770 0F 0M ro a/w/GKMUG lINVENTOR 04u/0 c2c/M5 K0 'F G.2.- BY

April 22, 1941. D. R. cLlMENKo l 2,238,973

SULPHONAMIDE COMPOUND YIN OIL Filed July 26, 1940 3 Sheets-Sheet 3 W C P m a M w W 4 H4 s4 HM l. M M @NAO oN @N40 mmf imfm?. V N 5 11H0 New 664m vw Emu im N 1.5mm s M C A 5F 0 4m E 4 M mw Ill 5 Maw Sm I CM m M .w W m. M M .w m-w 0C .M W. w .m my w w mv. m w O FIG.3.

Patented Apr.4 22, 1941 SULPHONAMIDE COMPOUND IN OIL David R. Climenko, Ellsmere, N. Y., assignor to American Cyanamid Company, New York, N. Y., a corporation of Maine Application July 26, 1940, Serial N0. 347,638

6 Claims.

This invention relates to chemotherapy and more particularly to methods and compositions for enhancing the therapeutic properties of sulphonamide compounds particularly for oral administration.

This application is in part a continuation of my application Serial No. 1939.

311,227 filed December 28,

in the therapeutic efvery marked improvement ,fect over the administration of an equivalent quantity of the sulphonamide compound or of compositions containing equivalent quantities of sulphonamide compounds To obtain the best results I prefer -to use an oil to drug ratio of from about 40:1 to 50:1.

It is an advantage of the present invention that a wide variety of fatty substances such as solid fats themselves and glyceride oils may be employed to produce the improved results. In fact, any fatty substance that is capable of being assimilated by the body will serve the purpose. The oils and fats are, with few exceptions, esters of the higher members of the fatty acids With the glyceryl radical and hence called glycerides of the fatty acids; therefore not only the oils and fats which are obtained from plant and animal tissue are useful in carrying out the present invention but also the various synthetic glycerides. Among the various fatty substances that have been found to be applicable to the present invention are lard, butter fat, corn oil, coconut oil, cottonseed oil, almond oil, peanut oil, olive oil, sesame oil, some fish oils, suet and synthetic triglycerides such as palmitin, olein, laurein and margarine. These fatty substances while they do e have value as foods peutically inactive.

It is also an advantage of the present invention that the therapeutic properties of all the active are considered to be therasulphonamide compounds are enhanced by administering along with the fatty substance which permits the selection of a particular sulphonamide compound in the treatment of the type of infection for which it has been found to 'be especially beneficial.

medium provided that certain minimum in an aqueous medium.

The sulphonamide compounds which have been found to be useful in carrying out the present invention may be represented by the following formula: (RSOZNRMX in which R is an amino or sulphonarnido aryl radical, R' is a radical included inthe group consisting of thiazoles, pyridines, pyrimidines, hydrogen, alkyl, aryl, or acyl radicals, and X is hydrogen or a metal and n is a small whole number.

Typical sulphonamide compounds useful in the invention include sulphanilamide itself, disulphanilamide, 2sulphanilamidothiazole, 2- sulphanilamidopyridine, 2sulphanilamidopyrim idine and the various Nl-acylsulphanilamides such as N 1-butyrylsulphanilamide and Nl-dodecanoylsulphanilamide and the like.

These compositions of the present invention are not to be confused With compositions in which sulphonamide compounds may have been administered in fatty mediums in which the proportion of oil to drug Was in the order of the ratio of about 4:1 or about a 20% concentration of the drug in oil. In these prior compositions, Which were usually administered by subcutaneous njections, there was no difference in the therapeutic effect produced than that which could have been obtained by administration of equivalent quantities of the sulphonamide compound in an aqueous medium and the c..l was usedonly for conveniencai. e., for the purpose of subcutaneous injection and to lessen chances for irritation due to the solid material at the site of the injection.

The enhanced therapeutic effects of the sulphonamide compounds when administered in a fatty medium have been proven by a number of experiments, the results of which are summarized in the following tables and are illustrated lby the drawings in which:

Fig. 1 is a graph which represents the therapeutic effects of sulphanilamide upon beta-hemo lytic streptococcal infections of mice when the compound is suspended in various amounts of olive oil, and illustrates the percentage mortality of the infected mice when suspensions of the drug in olive oil were administered in varying proportions;

Fig. 2 is a chart which represents the therapeutic effects of sulphanilamide upon beta-hemolytic streptococcal infections of mice when the compound is suspended in various amounts of olive oil, and illustrates the percentage average survival when suspensions of the drug in olive oil were administered in varying proportions;

Fig. 3 is a chart which represents the comparative therapeutic effects of various sulphonamide compounds in streptococcal infections of mice when the compounds are administered orally in acacia aqueous suspensions and in olive oil suspensions; and

Fig. 4 is a Chart which represents comparative 100:0. Each y and in the same experiment andeach performed with ,tions about 41 mice therapeutic effects of various sulphonamide compounds in pneumococcal infections of mice when the compounds are'administered orally -in aqueous acacia suspensions and in olive oil suspensions.

A numer of series of experiments were conducted to determine the comparative therapeutic effect of various combinations of olive oil and sulphanilamide when administered orally upon experimental beta-hemolytic. streptococcal infections in mice. In carrying out the experiments, suspensions were prepared of sulphanilamide in olive oil in which the ratio of oil to sulphanilamide were as follows: v50:1, 40:1, 30:1, 20:1, 10:1, 2:1,'0:1 (Water suspension of the drug), and of these suspensions was then tested for its therapeutic eiect upon the experimental infections in mice at the'saine time experiment was repeated a number of times. In each case Streptococcus strain C-203 was used which is a strain of beta-hemolytic Streptococcus well known inthe literature. The virulence of the strain was carefully checked by control animals all of which died. Administration of the drug was oral in each case, the amount being vings. per dose suspended in olive oil. The treatments were begun immediately after infection and repeated once daily for a total of three treatments. The mice were then observed for days and the average survival was computed on the basis of the survival for any part of the experimental period as well` as for the entire experimental period. The total number of animals was about '15 mice in the case of each of the varied proportions of oil and drug and the cumulative results agree in general with the results in the separate experiments done at various times on the smaller group of 25 mice. Data obtained in this fashion is usually considered more accurate than a single experiment 'Z5 mice. The results of the above tests are shown in the following table.

' This shows that the various quantities of I tality of the mice in about 5 mice out of 'I5 will survive when given 10 mg. oral doses of sulphanilamide in water. a mixture of olive oil and sulphanilamide, with oil to drug ratio being :1

5 is about ten times more effective than equivalent doses of sulphanilamide alone.

These results are graphically illustrated in Figs. 1 and 2. Fig. 2 indicates the percent average survival for the entire period of fteen days and is based upon the total number of mice used in the experiments. This chart shows that when the ratio of oil to sulphanilamide is only 2:1. or 10:1 there is very little improvement over the test using the drug alone (ratio 0:1). However, when the ratio of oil to drug is 20:1 the percent average survival is about 48% only about 25% survival when the same quantity of the drug is administered in an aqueous suspension. h Thel chart further shows that the percent average survival increases when the ratio of 30:1 isvused over that using 20:1. Itis further v noted that the maximum percent average survival which is about 69% is obtained when the ratio of 50:1 is used and that this was an improvement of only about 1V or 2% ratio indicating that while ratios of 50:1'or greater can be used without impairing the therapeutic effect, at the same time the improvement would not be much great r than that obtained using the 40:1 ratio. In Fig. 1 the percent morthe accumulated test is illustrated on the graph. The percent mortality is calculated for each of the days throughout the experiment. Here again there is shown to be very llittle difference between the therapeutic effect of the drug when administered in the ratio of 2:1 and 10:1 oil to drug and administration of an aqueous suspension or solution, while beginning with the 20: 1 ratio there is a definite and l marked improvement through each of the ratios up to the 50:1. The graph further shows that the rate of mortality is much greater in the case of the drug in aqueous suspension and. also when TAnLeNo. 1 The comparative therapeutic eects of various combinations of olive oil and stphanlamde when administered orally upon experimental beta-hemolytic streptococcal infection of mice y Deaths in days Survivals Ratio oil No t0 sul. Therapeutic mixture of Dose per os Per- Iliimi 1 2 4 s e 7 s 9 1o 11-15 No Per' ent cent average 2 gms. S. in 100 cc. oil. 50:1 75 10 mg. or 0.5 0c l 8 l2 10 35 46. 6 68. 5 2gms. S. in 807 oil in wateL-. 40:1 75 1 2 11 14 5 33 44.0 06.0 2 gms. S. in 60% oil in water.- 30:1 1 3 20 22 7 13 17.3 45. 9 2 gms. S. in 40% oil in Watan 20:1 13 7 27 12 4 1l 13.9 36.8 2 gms. S. in 20% oil m Water.. 10:1 13 26 22 8 5 4 5.0 27.0 2gms. S.in4% o1linwater 2:1 14 22 25 6 5 6.7 24.1 2 gms. S. in 100 oc. water 0:1 23 21 19 3 4 4 5.4 24.3 Olive oil controls 1:0 0 0 6.7 Controls 8 1 0 0 7.3

S.-Sulphanilamide.

`ci suaphanilamide are administered', whereas only vdata shows that after Jthe ratio of oil to drug is only about 10:1 than when the ratio is 20:1 orgreater and further, that at the end of five days substantially all of the kanimals have died that will die in the teen day period when the oil to drug ratio is 10W, Whereas a. much smaller number have died at the end of ve vdays when the larger ratios of oil to drug are used. This indicates that the infection is not only cured in a greater number of cases but also that the combination of the as compared to l over the 40:1 y

drug with larger quantities of oil in delaying the death of many Experiments the comparative therapeutic effect of various sulmediately after infection and repeated once daily The mice were for days and the average surthe results It is noted is a strain that is extremely susceptible therapeutic action.

to chemo- For this reason the drugs are considered therapeutically practical for pneumococcal infections. The generally enhanced therapeutic activity in the presence of glycerides is, however, maintained.

The reason Why the compositions of the present to be limited to any particular theory. It was thought that possibly the glycerides might produce increased or accelerated absorption of the administration of some other drugs with glycerides. .Accordingly the factor of absorption was inves igated by feeding mice 10 mg. doses of the compounds used in making the experiments shown apparent that with the exception of sulphanilamide itself there is no notable increase in absorption of the various drugs in olive oil as compared to suspensions in acacia. The compounds in these tests were also administered orally in 1/ cc. of olive oil or acacia solution.

TABLE 2 The blood concentration in mice obtained after oral administration of a series of NI-acyl derivatives of sulphanz'lamide Compound *E Blood levels in mg. percent hours following administration of the com- Dose Route pound Acacia p. o 20.0 10.9 5. 6 1. 0 T 0 Olive oil p. 0 34. 0 35. S 16.1 1l. 1 11. 1 3. 8 Acacia p. o 9.1 4. 0 1.1 T Olive oil p. o 8.6 4.5 2.6 0.6 Aca p. o 16. 6 10.0 2.9 0.6 Olive oil p. o 12. 5 11.5 7.1 2. 7 Acacia p. o 25.0 18.2 10.5 1. 7 Olive oil p. o 33.3 16.6 8.7 3.8 Acacia p o 18.8 16.6 12.5 1.9 l0 Olive oil p 18.0 13. 3 10.0 6. 2 10 cacla p. o 29.4 29.4 22. 2 9.0 10 Olive oil p. o 28.6 25.4 16.6 9. 1 l0 Acacia p o 30.0 28.6 16.2 T 10 Olive oil p. o 50.0 43.0 33. 3 21. 7 10 cia p o 20. 0 22.2 16.0 T

l0 35.3 30.0 16.0 16.0 l0 1l. 8 13. 8 6. 8 T 10 l2. 5 8. 9 6. 7 7. 3

+Alcohol precipitation method.

D. 0., per

T, trace.

FT, [aint trace.

with various compounds, being particularly no- ;iceable with the Nl-butyrylsulphanilamide.

Tests with other fats show similar results and .he enhanced therapeutic effect does not appear `o depend on the use of a. particular glyceride 1o long as it is readily assimilable by the animal Athat the amount of giyceride fat sion as the glyceride medium difference in absorption there is a very marked difference in therapeutic activity. In evaluating the results of the table which represents tests on a large number of animals, it should be noted that absorption varies quite markedly from animal to animal. The-figures in the table are averages but the variation which is normally noted in absorption is such that most of the differences are within the normal experimental error or normal variation. The therapeutic results on the other hand show much less variation so that the gures of the chart set out for Fig. 3 of the drawings are quite accurate.

It is an advantage of the present invention or oil used above a lcertain minimum proportion drug ratio is at least 20:1, is not critical and the ratio may be varied from 20-:1 up to 50:1 arid greater. Still larger amounts of fat do not appear to have any 'further useful effect. In general it may be stated that the increase in therapeutic activity will vary from drug to drug and it also varies to a lesser extent from glyceride to glyceride, and even from the glyceride in one physical form to the glyceride in a different physical form. In general, the fats which are rapidly assimilable ysuch as butter fats can be used in somewhat smaller amounts than is the case with fats which are very slightly assimilable such as certain very high melting fats. In every "case, however, the additions of the glyceride fat or oil enhance the therapeutic activity of the drugs as soon as it is present in substantial amounts. The activity rises to a maximum which will vary with the different drugs and further amounts of a glyceride do not show any particular beneficial action.

It is an advantage that the is not limited to any particular form of composition or method of administration. In many cases it is desirable to keep the drug in suspenappears to protect the drug from oxidation or other deterioration, panticularly when antioxidants are incorporated in the medium such as, ior example, thiourea and substituted pounds may also be administered in the form of oil and water emulsions provided.- the necessary oil to drug ratios are maintained.

The enhanced therapeutic activity of sulpresent invention l A may be taken advantage of in in which oil to y associated with glycerides either of two ways. The dosage of drug may be maintained constant and an improved therapeutic effect obtained, or a smaller dosage may be used to obtain the same therapeutic effect. Of course, methods combining these two may be used and in general the form in which the compounds of the present invention are utilized will depend on whether in a particular case it is more important to reduce toxic effects incident to iarge dosagesy or to obtain even greater therapeutic activity;y It is thus an advantage of the present invention that it is very exible and the treatment calculated to give most effective and safest resul-ts in any given casev may be chosen.

The enhanced therapeutic eiect of the present invention is obtained when the drug is introduced through the alimentary system either by oral or rectal administration. The compositions of the present invention are not, in generaLfsuitable for parenteral administration.

What I claim is:

1. A therapeutic composition for oral administration which comprises a sulphonamide compound having therapeutic properties and a Afatty substance of the group consisting and glycerides contained therein, in which the ratio of the fatty substance to the sulphonamide compound is at least 20:1.

2. A therapeutic composition for oral administration which comprises a sulphonamde compound having therapeutic properties and a fatty substance of the groupconsisting of oils, fats, and glycerides. contained therein, in which the ratio of the fatty substance to the sulphonamide compound is from about 40:1 to 50:1.

3. A composition according to claim 1 in which the sulphonamide compound is sulphanilamide.

4. A composition according to claim 1 in which the sulphonamide compound is -an Nl-acylsulphanilamide.

5. A composition according to claim 1 in which phonamide drugs when the sulphonamde compound is Nl-butyrylsulthioureas. The comf phanilamide.

6. A composition according to claim 1 in which the sulphonamide compound is Nl-dodecanoylsulphanilamide.

DAVID R.. CLIMENKO.

of oils, fats,` 

